Breast cancer is among the most prevalent forms of cancer worldwide, representing a significant cause of morbidity and mortality. This accounts for approximately 25 % of all cancer cases in women, with an annual incidence rate that persists in the upward trajectory (Arnold et al., 2022). Despite significant advancements in detection and treatment, breast cancer continues to represent a substantial challenge in the field of oncology, showing the necessity for the development of more effective and specific interventions(G.Waks and Winer, 2019; Obeagu and Obeagu, 2024). The Michigan Cancer Foundation-7 (MCF-7) breast cancer cell line is a model used extensively in cancer investigations because of the hormone-responsive characteristics, with approximately 70 % of cases in women being associated with sensitivity to estrogen (Barrios, 2022; W. Cao et al., 2021). The MCF-7 cell line, which originates from human breast adenocarcinoma, is a model widely used in evaluating treatment responses to anticancer agents, particularly those modulating the estrogen pathway (Tran et al., 2021; Wang et al., 2021a, Wang et al., 2021b). In ER-positive breast cancer, BCL-2 is a key pro-survival node downstream of estrogen signaling. This ER→BCL-2 axis is clinically relevant and has been explored with venetoclax in ER+/BCL-2+ disease. Therefore, a decrease in BCL-2 is biologically meaningful in ER + MCF-7 when survival signaling is suppressed (Kawiak and Kostecka, 2022; Lindeman et al., 2022; Lok et al., 2019; Mehraj et al., 2024).

Primary treatment interventions for breast cancer include surgical procedures, chemotherapy, and radiotherapy. These have been proven to reduce tumor size and prevent metastasis, but are associated with substantial limitations, such as the occurrence of toxic side effects, drug resistance development, and the potential for damage to healthy tissue(Abbas and Alrumman, 2022; Pe et al., 2022). Consequently, there has been a surge of interest in natural compound-based methods, which offer a reduced toxicity profile and the capacity to target specific molecular pathways in cancer cells (Koual et al., 2020).

In Indonesia, traditional medicine has long relied on a wide variety of medicinal plants for the treatment of major diseases, including cancer. Among these, the root of Poikilospermum suaveolens (Blume) Merr. (P. suaveolens), locally known as "bajakah," has been popularly used by indigenous communities, especially in Kalimantan and Sulawesi, for generations as a herbal remedy for cancer, wound healing, and general health (Afni Bandy et al., 2021; Hapid et al., 2021b; Hartati et al., 2022). This ethnomedicinal relevance is supported by several surveys reporting its traditional use among Dayak communities in Borneo and Sulawesi. The development of natural compound-based anticancer treatment may be facilitated by P. suaveolens, a member of the Urticaceae family that has a long history in traditional medicine. Previous studies reported that the ethyl acetate fraction of P. suaveolens (FEAPs) exerted anticancer activity through mechanisms including apoptosis and cell proliferation inhibition (Rooney et al., 2021). FEAPs has been found to contain a variety of bioactive compounds, including flavonoids, saponins, tannins, and triterpenoids, which have the potential to act as anticancer, anti-inflammatory, antioxidant, and antimicrobial agents (Hartati et al., 2020; Sattar et al., 2023). This traditional use has been supported by several ethnobotanical surveys and is consistent with the reported anticancer properties of other Urticaceae family members.

As reported in previous studies on plants belonging to the Urticaceae family, including Urtica dioica and Urtica urens, there is compelling evidence of anticancer properties, characterized by cell proliferation inhibition and apoptosis promotion(Bandy et al., 2021; Jumania et al., 2020). Urtica dioica extract is established to be capable of increasing the expression of both caspase-3 (CASP-3) and caspase-9 (CASP-9). This observation suggests the activation of the apoptotic pathway, which plays a crucial role in the elimination of cancer cells (Wright, 2018). Notably, MCF-7 is CASP-3 deficient due to a 47-bp deletion in exon 3, thus CASP-3 is typically undetectable in this model. The absence of CASP-3 bands should be interpreted as a cell-line characteristic, not a treatment effect (Janicke, 2009; Tian, 2023; S. Wang et al., 2016).

Molecular docking has become a significant computational instrument in the field of drug discovery. The instrument allows the prediction and analysis of the interactions between bioactive compounds and specific molecular targets in cancer cells (Ahmad et al., 2024; Bahadur et al., 2021; Siddiqui et al., 2021). In this study, docking is used exploratorily as a hypothesis-generating approach. Given the casp-3 deficiency of MCF-7, any casp-3 docking is treated as comparative only (structure-based benchmarking), not as evidence of functional activity in this cell line. Our functional focus is on casp-7/-8/-9, PARP-1, and BCL-2, which align with ER + biology and apoptosis regulation in MCF-7 (Mutakin et al., 2025). Previous molecular docking studies reported that various flavonoids and phenolic compounds such as quercetin and taxifolin could bind effectively with Casp-3, showing the role played in modulating the apoptotic signaling pathway. Molecular docking studies are used to determine specific molecular interactions supporting the anticancer properties of natural compounds.

The potential of P. suaveolens to inhibit the proliferation of breast cancer cells was suggested based on the ability reported by the Urticaceae family. Despite its popularity in traditional medicine, the scientific validation and mechanistic evidence supporting the anticancer effects of P. suaveolens roots are still limited. Thus, a rigorous pharmacological investigation is required to bridge traditional claims with modern biomedical science. Therefore, this study aimed to explore the molecular mechanisms of the anticancer activity of FEAPs against MCF-7 cells. The applied methods included phytochemical analysis using Liquid Chromatography-High Resolution Mass Spectrometry/Tandem Mass Spectrometry (LC-HRMS/MS) to determine the major bioactive compounds. Additionally, the IC50 value was determined through cytotoxic tests, followed by morphological observations of cancer cells. Molecular docking was conducted as a hypothesis-generating tool to explore interactions with apoptosis and survival targets relevant to ER + MCF-7 (e.g., Casp-7/8/9, PARP-1, BCL-2, casp-3 included comparatively). This design links our LC-HRMS/MS profile, cytotoxicity, morphology, and transcript changes to a coherent mechanism consistent with CASP-3 deficient MCF-7 and supports development of targeted, natural compound based anticancer strategies.